| 摘 要： | Objective: To investigate the mechanism of c AMP-PKA signaling pathway mediated by Chinese medicine formula Shaoyao Gancao Decoction(芍药甘草汤, SGD) on the regulation of aquaporin 5(AQP5) and muscarinic receptor 3(M3 R) levels in Sj?gren's syndrome(SS). Methods: Of the 30 mice, 5 were randomly selected as control, and others were used for creating SS model. After successful modeling, mice were randomly divided into 5 groups(n=5 per group) and intragastrical y administered with saline(8 m L/kg), pilocarpine(1.4 mg/kg), or low, medium and high doses SGD(0.14, 0.21, 0.35 g/kg Radix paeoniae with 0.01 g/kg Radix glycyrrhizae, respectively) for 6 weeks. Human labial gland acinar cel s were treated with pilocarpine or varying doses of SGD with saline as the placebo. Hematoxylin and eosin staining was used to observe the histopathological changes of the submandibular glands of mice. The serum levels of anti-SS antigen A(SS-A), anti-SS antigen B(SS-B), M3 R, and α-fodrin in submandibular glands of mice were measured by enzyme-linked immunosorbent assay. Immunofluorescence staining was used to observe the spatial localization of AQP5 and M3 R in acinar cells. Reverse transcriptase polymerase chain reaction and Western blot were used to detect the expressions of PKA, c AMP, Epac1, AQP5, M3 R, nuclear factor kappa-B(NF-κB), and tumor necrosis factor(TNF)-α in submandibular gland tissues and cel s of each group. Results: Compared to normal mice, body weight, 5-min salivary secretion, 30-min secretion of tears and breakup time of tear film of model mice decreased at 1–6 weeks after immunization(al P0.05), whereas water intake increased(al P0.05). In the model group, glands of the submandibular glands showed atrophy, accompanied by acini of different sizes, decreased numbers and loose arrangement, with catheter dilatation and different degrees of lymphocyte infiltration. Conditions of mice in SGD groups were improved. The positive expression of AQP5 and M3 R were higher in the acinar cel s treated with al doses SGD compared to the normal group; serum levels of SS-A, SS-B, and α-fodrin were lower, and that of M3 R was higher in al doses SGD treated animals than the model or pilocarpine treated ones(al P0.05). Compared to the model and pilocarpine groups, the m RNA and protein levels of NF-κB and TNF-α were lower in mice or cel s treated with medium or high-dose SGD(al P0.05), while those of PKA, Epac1, AQP5 and M3 R were higher(al P0.05). Conclusion: SGD can improve symptoms of SS by regulating the c AMP-PKA signaling pathway and increasing AQP5 and M3 R levels.
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